MicroRNAs (miRNAs) are important regulators of plant growth and development. Previously, we identified a group of conserved and novel miRNA families from soybean (Glycine max) roots. Many of these miRNAs are specifically induced during soybean-Bradyrhizobium japonicum interactions. Here, we examined the gene expression levels of six families of novel miRNAs and investigated their functions in nodule development. We used northern-blot analyses to study the tissue specificity and time course of miRNA expression. Transgenic expression of miR482, miR1512, and miR1515 led to significant increases of nodule numbers, while root length, lateral root density, and the number of nodule primordia were not altered in all tested miRNA lines. We also found differential expression of these miRNAs in nonnodulating and supernodulating soybean mutants. The expression levels of 22 predicted target genes regulated by six novel miRNAs were studied by real-time polymerase chain reaction and quantitative real-time polymerase chain reaction. These results suggested that miRNAs play important roles in soybean nodule development.
Flavones, a major group of flavonoids in most plant tissues, play multiple roles in plant-environment interactions. In our study, the expression of the two soybean flavone synthase genes, GmFNSII-1 and GmFNSII-2, was significantly increased by methyl jasmonate (MeJA), glucose, mannitol and NaCl treatment, which were also found to increase flavone aglycone accumulation in Glycine max (L.) Merrill. In the GmFNSII-1 promoter, a specific CGTCA motif in the region (-979 bp to -806 bp) involved in the MeJA response was identified. Promoter deletion analysis of GmFNSII-2 revealed the presence of osmotic-responsive (-1,143 bp to -767 bp) and glucose-repressive sequence elements (-767 bp to -475 bp), which strongly supported the hypothesis that glucose induces soybean flavone production by acting as both an osmotic factor and a sugar signaling molecule simultaneously. Silencing of the GmFNSII gene clearly reduced the production of flavone aglycones (apigenin, luteolin and 7,4'-dihydroxyflavone) in hairy roots. The GmFNSII-RNAi (RNA interference) roots that had a reduced level of flavones accompanied by more malondialdehyde and H2O2 accumulation were more sensitive to salt stress compared with those of the control, and we concluded that flavones, as antioxidants, are associated with salt tolerance.
Soybean flavonoids, a group of important signaling molecules in plant-environment interaction, ubiquitously exist in soybean and are tightly regulated by many genes. Here we reported that GmMYB100, a gene encoding a R2R3 MYB transcription factor, is involved in soybean flavonoid biosynthesis. GmMYB100 is mainly expressed in flowers, leaves and immature embryo, and its level is decreased after pod ripening. Subcellular localization assay indicates that GmMYB100 is a nuclear protein. GmMYB100 has transactivation ability revealed by a yeast functional assay; whereas bioinformatic analysis suggests that GmMYB100 has a negative function in flavonoid biosynthesis. GmMYB100-overexpression represses the transcript levels of flavonoid-related genes in transgenic soybean hairy roots and Arabidopsis, and inhibits isoflavonoid (soybean) and flavonol (Arabidopsis) production in transgenic plants. Furthermore, the transcript levels of six flavonoid-related genes and flavonoid (isoflavonoid and flavone aglycones) accumulation are elevated in the GmMYB100-RNAi transgenic hairy roots. We also demonstrate that GmMYB100 protein depresses the promoter activities of soybean chalcone synthase and chalcone isomerase. These findings indicate that GmMYB100 is a negative regulator in soybean flavonoid biosynthesis pathway.
Priming can improve plant innate capability to deal with the stresses caused by both biotic and abiotic factors. In this study, the effect of DL-β-amino-n-butyric acid (BABA) against Aphis glycines Matsumura, the soybean aphid (SA) was evaluated. We found that 25 mM BABA as a root drench had minimal adverse impact on plant growth and also efficiently protected soybean from SA infestation. In both choice and non-choice tests, SA number was significantly decreased to a low level in soybean seedlings drenched with 25 mM BABA compared to the control counterparts. BABA treatment resulted in a significant increase in the activities of several defense enzymes, such as phenylalanine ammonia-lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO), chitinase (CHI), and β-1, 3-glucanase (GLU) in soybean seedlings attacked by aphid. Meanwhile, the induction of 15 defense-related genes by aphid, such as AOS, CHS, MMP2, NPR1-1, NPR1-2, and PR genes, were significantly augmented in BABA-treated soybean seedlings. Our study suggest that BABA application is a promising way to enhance soybean resistance against SA.
Inspired from falling leaves, ZnO nanorods-nanoslices hierarchical structure (NHS) was constructed to modify the surfaces of two widely used implant materials: titanium (Ti) and tantalum (Ta), respectively. By which means, two-stage release of antibacterial active substances were realized to address the clinical importance of long-term broad-spectrum antibacterial activity. At early stages (within 48 h), the NHS exhibited a rapid releasing to kill the bacteria around the implant immediately. At a second stage (over 2 weeks), the NHS exhibited a slow releasing to realize long-term inhibition. The excellent antibacterial activity of ZnO NHS was confirmed once again by animal test in vivo. According to the subsequent experiments, the ZnO NHS coating exhibited the great advantage of high efficiency, low toxicity, and long-term durability, which could be a feasible manner to prevent the abuse of antibiotics on implant-related surgery.
Previous reports indicated that AtMYB103 has an important role in tapetum development, callose dissolution, and exine formation in A. thaliana anthers. Here, we further characterized its function in anther development by expression pattern analysis, transmission electron microscopy observation of the knockout mutant, and microarray analysis of downstream genes. A total of 818 genes differentially expressed between ms188 and the wild-type were identified by global expression profiling analysis. Functional classification showed that loss-of-function of AtMYB103 impairs cell wall modification, lipid metabolic pathways, and signal transduction throughout anther development. RNA in situ hybridization confirmed that transcription factors acting downstream of AtMYB103 (At1g06280 and At1g02040) were expressed in the tapetum and microspores at later stages, suggesting that they might have important roles in microsporogenesis. These results indicated that AtMYB103 is a crucial regulator of Arabidopsis anther development.
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